生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
48 巻, 1 号
選択された号の論文の8件中1~8を表示しています
  • 中北 愼一, 長束 俊治, 池中 一裕, 長谷 純宏
    2004 年 48 巻 1 号 p. 1-4
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    我々は以前, マウスの各臓器からN-配糖体糖鎖を調製し, 逆相HPLCによって比較分析を行った. その結果, 中枢神経系に特異的に発現している2種類の糖鎖 (BA-1, BA-2) を見出し, それらの構造を決定した. 特にBA-2は相対量が多く, 大脳の全N-配糖体糖鎖量の5%, 小脳では7%を占めていた. また, マウスの脳の発達初期段階において強く発現しており, 神経突起の伸長やシナプスの形成に関与しているのではないかと推察されている. 今回我々は, BA-1, BA-2がマウスの脳における糖鎖の生合成過程での中間産物ではなく, 構造的にも突出した糖鎖であり, その生合成には非常に厳密な基質認識を行う“脳型”ガラクトース転移酵素が重要であることについて, ピリジルアミノ化法を用いた糖鎖構造解析とガラクトース転移酵素の基質特異性を解析することで明らかにした.
  • 川崎 ナナ, 橋井 則貴, 伊藤 さつき, 日向 昌司, 川西 徹, 早川 堯夫
    2004 年 48 巻 1 号 p. 5-10
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    Liquid chromatography/mass spectrometry equipped with a graphitized carbon column is useful for the simultaneous analysis of oligosaccharides. By using capillary column and nanoelectrospray ion source, the method can be used for the oligosaccharide profiling of sub microgram quantities of glycoproteins. This oligosaccharide profiling is expected to be a powerful tool for the glycome analysis. We demonstrate a potential application of oligosaccharide profiling in glycomics with two examples, the structural analysis of N-linked oligosaccharides from a gel-separated glycoprotein, and the differential analysis of N-linked oligosaccharides in cells.
  • 平林 淳
    2004 年 48 巻 1 号 p. 11-17
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    “Functional glycomics”is a coming area of glycoscience and glycotechnology, which aims at elucidating biological roles of protein glycosylation from a holistic viewpoint, in a particular context of post-genomics and post-proteomics. However, analysis of glycans, i.e., the third bio-informative chain, should meet various difficulties due to many factors which other bio-macromolecules lack, e.g., branching and linkage isomerism. Recently, unique methods, which enable a large scale of identification methods of glycoproteins have been developed independently in two groups. In this chapter, advanced methods for glycoproteomics enabling identification of both core proteins and glycosylation sites are described as well as subsequent glycan profiling by means of lectin-affinity technologies.
  • 西原 祥子
    2004 年 48 巻 1 号 p. 19-24
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    Elucidation of the biological role of glycan is one of the most important subjects to be resolved following the genome project. Glycosylation of proteins and lipids is performed in the Golgi apparatus by glycosyltransferases, which are responsible for synthesizing the huge diversity of complex oligosaccharides attached to glycoproteins and glycolipids. Our molecular evolutionary study showed that a prototype of each glycosyltransferase was conserved in Drosophila, suggesting common roles of glycans in humans and Drosophila.
    RNA interference (RNAi) was first reported in 1998 as a biological response of C. elegans to exogeneous double-strand RNA (dsRNA), which induces sequence-specific gene silencing. It is a multi-step process including the generation of active small interfering RNA (siRNA) by reaction with an RNase III endonuclease, Dicer. The resulting 21- to 23-nt siRNA mediates degeneration of the complementary homologous RNA. RNAi has recently emerged as a powerful reverse genetics tool for studying gene function in many model organisms, including Drosophila.
    To analyze the basic physiological functions of glycans, we established the Drosophila RNAi knock down system of glycosyltransferases and verified the system using the Drosophila proteoglycan UDP-galactose: β-xylose β1, 4galactosyltransferase I (dβ4GalTI). The expression of the target gene was disrupted specifically and the degree of interference was correlated with the phenotype. This study was the first to use reverse genetics, RNA interference, to study Drosophila glycosyltransferase systematically. The inducible glycosyltransferase RNAi knock down fly obtained using the GAL4-UAS system will open a new way for the analysis of the biological role of glycans.
  • 垣沼 直人, 佐藤 正明, 山田 達也, 河府 和義, 中島 元夫, 秋山 徹, 大和田 進, 芝中 安彦
    2004 年 48 巻 1 号 p. 25-30
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    In-gel competitive reassociation (IGCR) technique is one of the DNA subtraction methods with a high capacity. We have improved the IGCR technique and overcame several disadvantages such as time-consuming and complex process with poor reproducibility. The improved IGCR method was applied to the analysis of human gastric adenocarcinoma genomic DNA. The genomic DNA library, which was constructed after the subtraction by IGCR of a normal gastric tissue genomic DNA from an adenocarcinoma genomic DNA in the same patient, contained 13.9% clones of Epstein-Barr virus (EBV) genomic DNA fragment, which is known to be a cause of Burkitt lymphoma. The quantification of EBV genomic DNA revealed that the detection of these EBV fragments originated in the gastric adenocarcinoma genomic DNA was due to the condensation from a few copies to about 100 thousand copies by the improved IGCR technique. This suggests that our improved IGCR technique can efficiently enrich differences between two genomic DNAs with high simplicity.
  • 増田 豪, 石井 奈々, 君島 香奈子, 天野 卓, 横濱 道成
    2004 年 48 巻 1 号 p. 31-36
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    ヤギ血清中のALPには小腸, 骨および肝臓由来の3種類が確認された. 由来臓器型は電気泳動パターンから, 5タイプに分類した. また, SGE法で検出されるF型バンドは肝臓由来のALPだった. AlkPhor SYSTEMを用いることでSGE法によるF型と0/0型はさらにそれぞれF/FF/0型およびF/00/0型に分類できた. ALP由来臓器は加齢によって変化した. 骨由来のALPのみで構成される個体(0/0型)は3歳齢以降で確認されなかったことから, ヤギの体高および体長に関わる基本的な骨格形成は3歳齢までにほぼ終了するものと考えられた. 肝型ALPを発現しない0/0型の個体は3歳齢までに死亡していた. 韓国在来ヤギ集団はALP活性値から高活性値群(500IU/l以上)と低活性値群(500IU/l未満)の2群にわけられ, シバヤギにない特性が認められた.
  • 新井 博文, 加藤 陽二, 福永 憲次, 毛利 哲, 中村 和行
    2004 年 48 巻 1 号 p. 37-40
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    A mechanism of oxidative modification of apolipoproteins (apo) in human very-low-density lipoprotein (VLDL) was investigated in vitro. Lipid peroxidation was promoted by cupric ion in VLDL. Modification of apoE and apoB-100 was observed in the VLDL oxidation. Nε-(Hexanonyl)lysine, one of the lipid hydroperoxide-modified lysine residue, was detected in VLDL oxidized for 18 hours by immunoblot analysis and enzyme-linked immunosorbent assay. The results indicate that lysine residues of apoE and apoB-100 were modified by lipid hydroperoxides. The heparin-binding activity of apoE and apoB-100 which seems to reflect their low-density lipoprotein receptor (LDLr)-binding activity decreased in the VLDL oxidation. This demonstrates that the heparin-binding site of apoE and apoB-100 which includes lysine residues was modified in the VLDL oxidation. Our data suggest that lysine residues of the LDLr-binding site of apoE and apoB-100 might be damaged by lipid hydroperoxides produced in the VLDL oxidation.
  • 大谷 絵美, 平山 博樹, 横濱 道成
    2004 年 48 巻 1 号 p. 41-43
    発行日: 2004/03/15
    公開日: 2009/03/31
    ジャーナル フリー
    Cattle milk proteins are composed of casein components (Cn) such as α, β and κ-Cn, and whey components such as β-lactoglobulin (Lg). These milk protein components were separated by two-dimensional electrophoresis and compared with the separation patterns of yak, goat, sheep, deer, camel, llama, pig, horse and human milk. Based on our results, cattle and yak, goat and sheep, and camel and llama had very similar patterns, respectively.
    As for components like β-Lg in deer, were separated at the same location as that of cattle, components like α- and β-Cn resembled the patterns of goat and sheep. Also, the separation patterns of κ-Cn were divided into two groups: animals (cattle, yak, goat, sheep and llama) whose milk separated near pI6.3, and animals (deer, horse and human) whose milk separated near pI6.9.
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